Glycogen-depleted State Research Articles

Intake Of Essential Amino Acids Stimulates Mtorc1 Signaling And Inhibits Autophagy Following Glycogen-depleted Resistance Exercise

Autophagy is responsible for degrading and recycling cellular proteins under conditions of energetic stress and low nutrient availability. Activation of autophagy is mediated by AMPK which is activated by high-intensity exercise and low glycogen availability and performing resistance exercise with low glycogen levels may result in enhanced autophagy activation which in turn may have negative effects on muscle protein balance. Autophagy is inhibited by mTORC1 which is potently activated by essential amino acids (EAA). The opposing effects of AMPK and mTORC1 on autophagy suggests that any potential negative effects of performing resistance exercise with low glycogen availability may be rescued by EAA intake. PURPOSE: To study the effect of EAA intake on autophagy and mTORC1 signaling following resistance exercise with high and low glycogen availability. METHODS: Using a unilateral study design in which one leg was glycogen loaded and the other leg was glycogen depleted, men and women underwent two experimental trials wherein they consumed either a placebo (PLA) or an EAA drink after resistance exercise. Unilateral differences in muscle glycogen content were achieved through glycogen loading and subsequent one-legged glycogen depleting exercise the evening prior to each experimental trial. Muscle biopsies were collected at baseline, post exercise and 1 and 3 h after drink intake in both legs in both trials. RESULTS: Mean glycogen content was ~69 % lower in the depleted leg compared to the loaded leg (228 vs 724 mmol/kg dry weight) across all time points in both trials. Phosphorylation of ULK1 at the AMPK-specific site S317 was elevated ~5-fold immediately post exercise in the depleted leg while remaining unchanged in the loaded leg with no differences between trials. In the PLA-trial, S317 phosphorylation remained elevated 60 min post drink ingestion, while in the EAA trial at the same time point, it had returned to baseline values. Phosphorylation of S6K1 at the mTORC1-specific site T389 remained largely unchanged at all time-points in the PLA-trial while being increased by ~33-fold in both legs at 60 min in the EAA-trial. CONCLUSIONS: Intake of EAA blunt the elevations in autophagy signaling induced by resistance exercise performed in a glycogen-depleted state while simultaneously stimulating the mTORC1 pathway.

Endogenous glucose production increases in response to metformin treatment in the glycogen-depleted state in humans: a randomised trial.

Metformin is believed to reduce glucose levels primarily by inhibiting hepatic glucose production. Recent data indicate that metformin antagonises glucagon-dependent glucose output, suggesting that compensatory mechanisms protect against hypoglycaemia. Here, we examined the effect of metformin on glucose metabolism in humans after a glycogen-depleting fast and the role of reduced-function alleles in OCT1 (also known as SLC22A1). In a randomised, crossover trial, healthy individuals with or without reduced-function alleles in OCT1 were fasted for 42 h twice, either with or without prior treatment with 1 g metformin twice daily. Participants were recruited from the Pharmacogenomics Biobank of the University of Southern Denmark. Treatment allocation was generated by the Good Clinical Practice Unit, Odense University Hospital, Denmark. Variables of whole-body glucose metabolism were assessed using [3-(3)H]glucose, indirect calorimetry and measurement of substrates and counter-regulatory hormones. The primary outcome was endogenous glucose production (EGP). Thirty-seven individuals were randomised. Thirty-four completed the study (12 had none, 13 had one and nine had two reduced-function alleles in OCT1). Three were excluded from the analysis because of early dropout. Metformin significantly stimulated glucose disposal rates and non-oxidative glucose metabolism with no effect on glucose oxidation. This increase in glucose utilisation was explained by a concomitant increase in glycolytic flux and accompanied by increased EGP, most likely mediated by increased plasma lactate, glucagon and cortisol levels. There was no effect of reduced-function OCT1 alleles on any of these measures. All individuals completed the glycogen-depleting fast without hypoglycaemia. Metformin stimulates glycolytic glucose utilisation and lactate production in the glycogen-depleted state. This may trigger a rise in glucose counter-regulatory hormones and subsequently an increase in EGP, which protects against hypoglycaemia. ClinicalTrials.gov NCT01400191 FUNDING: Danish Research Council for Health and Disease (0602-02695B) and Odense University Hospital Free Research Fund, 2012.

Protein ingestion does not impair exercise-induced AMPK signalling when in a glycogen-depleted state: implications for train-low compete-high

The aim of the present study was to test the hypothesis that consuming protein does not attenuate AMPK signalling when exercise is commenced in a glycogen-depleted state. After performing a glycogen-depleting protocol the evening before, the subsequent morning ten active men performed 45 min steady-state cycling at 50 % of peak power output (PPO) followed by an exercise capacity test (1-min intervals at 80 % PPO interspersed with 1-min periods at 40 % PPO). In a repeated measures design, subjects consumed 20 g of a casein hydrolysate solution (PRO) 45 min before exercise, 10 g during and a further 20 g immediately post-exercise, or an equivalent volume of a non-calorie taste matched placebo (PLA). Resting (PRO = 134 ± 29; PLA = 136 ± 28 mmol kg(-1)) and post-exercise muscle glycogen (PRO = 43 ± 16; PLA = 47 ± 18 mmol kg(-1)) was not different (P > 0.05) between trials nor was exercise capacity (PRO = 26 ± 9; PLA = 25 ± 10 min, P > 0.05). Phosphorylation of AMPK(Thr172) increased threefold immediately post-exercise (P < 0.05) and PGC1-mRNA increased sixfold at 3 h post-exercise (P < 0.05), though there were no differences between conditions (P > 0.05). In contrast, there was a trend (P = 0.08) for a divergent response in eEF2(Thr56) phosphorylation such that 1.5 fold increases post- and 3 h post-exercise in PLA were blunted with PRO, thus indicative of greater eEF2 activation. We conclude that athletes who deliberately incorporate training phases with reduced muscle glycogen into their training programmes may consume protein before, during and after exercise without negating signalling through the AMPK cascade.

Glycogen depletion increases peroxisome proliferator activated receptor‐δ (PPAR‐δ) activity following acute exercise

Performing exercise with low muscle glycogen has profound effects on substrate utilization and gene transcription. The aim of the present study was to examine signaling pathways potentially responsible for modulating gene transcription during exercise in a glycogen-depleted state. Methods Twenty female Wistar rats were randomly assigned to 5 experimental groups: control [CON]; normal glycogen control [NG-C]; normal glycogen exercise [NG-E]; low glycogen control [LG-C]; and low glycogen exercise [LG-E]). Results Exercise reduced muscle glycogen by 47 and 75% post exercise in the NG-E and LG-E groups respectively. The exercise protocol meant that the LG-E group began exercise with a 40% decrease in resting glycogen levels (2.1 ± 0.1 to 1.3 ± 0.3 nmol.mg.wet wt; P<0.05). Exercise in low glycogen increased AMPK α1 activity by 32% (CON 0.04 ± 0.002, LG-E 0.06 ± 0.006 Units mg/AMPK; P<0.05) and AMPK α2 activity by 147% (CON 0.03 ± 0.004, LG-E 0.07 ± 0.005 Units mg/AMPK; P<0.05). In addition to the increase in AMPK activity, ChIP analysis demonstrated increased PPAR-δ binding to the CPT1 promoter in the LG-C, LG-E, and NG-E experimental groups compared to the CON group or the NG-C groups. Discussion Our data suggests that PPAR-δ activation may contribute to the altered gene transcription previously reported during exercise in a glycogen-depleted state.

Reliability of a cycling time trial in a glycogen-depleted state

The aim of this study was to investigate the reliability of a protocol designed to simulate endurance performance in events of long duration (approximately 5 h) where endogenous carbohydrate stores are low. Seven male subjects were recruited (age 27 +/- 7 years, VO(2max) 66 +/- 5 ml/kg/min, W (max) 367 +/- 42 W). The subjects underwent three trials to determine the reliability of the protocol. For each trial subjects entered the laboratory in the evening to undergo a glycogen-depleting exercise trial lasting approximately 2.5 h. The subjects returned the following morning in a fasted state to undertake a 1-h steady-state ride at 50% W (max) followed by a time trial of approximately 40-min duration. Each trial was separated by 7-14 days. The trials were analysed for reliability of time to completion of the time trial using a coefficient of variation (CV), with 95% confidence intervals (data are mean +/- SD). The times to complete the three trials were 2,546 +/- 529, 2,585 +/- 490 and 2,568 +/- 555 s for trials 1, 2 and 3, respectively. The CV between trials 1 and 2 was 4.5% (95% CI 2.9-10.4%) and between trials 2 and 3, 3.8% (95% CI 2.4-9.9%). There was no difference in oxygen uptake, respiratory exchange ratio, carbohydrate oxidation, fat oxidation, plasma glucose concentration and plasma lactate concentration between the three trials. Therefore we can conclude that prior glycogen depletion does produce a reliable measure of performance with metabolic characteristics similar to ultraendurance exercise.

Regulation of muscle glucose transport during exercise.

In the present short review some factors affecting glucose utilization during exercise in skeletal muscle will be briefly described. Special focus will be put on the glucose transport step across the sarcolemma. Glucose transporters (GLUT4) are expressed at a surprisingly similar level in the different muscle fiber types in human skeletal muscle in contrast to findings in the rat. When working at the same absolute work load muscle glucose transport is decreased in trained compared with untrained muscle in part due to a decrease in GLUT4 translocation to the sarcolemma in trained muscle. However, when trained and untrained muscle are stressed severely by a workload taxing 100% of their peak oxygen uptake in a glycogen-depleted state, then glucose uptake is larger in trained than in untrained muscle and correlates with muscle GLUT4 content. Finally, the possible role of the AMP-activated protein kinase (AMPK) in regulating glucose uptake during exercise is discussed. It is indicated that at present no experiments definitively link activation of AMPK to activation of muscle glucose transport during exercise.

Glucose uptake is increased in trained vs. untrained muscle during heavy exercise.

Endurance training increases muscle content of glucose transporter proteins (GLUT-4) but decreases glucose utilization during exercise at a given absolute submaximal intensity. We hypothesized that glucose uptake might be higher in trained vs. untrained muscle during heavy exercise in the glycogen-depleted state. Eight untrained subjects endurance trained one thigh for 3 wk using a knee-extensor ergometer. The subjects then performed two-legged glycogen-depleting exercise and consumed a carbohydrate-free meal thereafter to keep muscle glycogen concentration low. The next morning, subjects performed dynamic knee extensions with both thighs simultaneously at 60, 80, and until exhaustion at 100% of each thigh's peak workload. Glucose uptake was similar in both thighs during exercise at 60% of thigh peak workload. At the end of 80 and at 100% of peak workload, glucose uptake was on average 33 and 22% higher, respectively, in trained compared with untrained muscle (P < 0.05). Training increased the muscle content of GLUT-4 by 66% (P < 0. 05). At exhaustion, glucose extraction correlated significantly (r = 0.61) with total muscle GLUT-4 protein. Thus, when working at a high load with low glycogen concentrations, muscle glucose uptake is significantly higher in trained than in untrained muscle. This may be due to the higher GLUT-4 protein concentration in trained muscle.

16 METABOLIC RESPONSE TO MODERATE EXERCISE IN LAMBS WITH AN AORTOPULMONARY SHUNT

The normal metabolic response to moderate exercise consists of a slight increase in glucose (glue) and a considerable increase in free fatty acids (FFA) in blood. In earlier studies we have demonstrated that at rest, after an overnight fast, lambs with an aortopulmonary shunt (SH) had lower concentrations of glue and FFA than control (C) lambs. We wondered, whether SH lambs with low glue and FFA were able to increase their arterial concentrations during exercise just like C lambs. Therefore, we studied 6 7-week-old SH lambs and 6 C lambs of the same age after an overnight fast at rest and during moderate exercise (tredmill; 50 % of Vo2-max; 30 rain). At rest as well as during exercise, 3 blood samples were taken at intervals of 10 min. At rest, mean arterial concentrations (mmol/l) of glue (SH: 3.37 ± 0.21 vs. C; 4.48 ± 0.53, mean ± SD, p < 0.05) and FFA (SH: 0.57 ± 0.17 vs. C: 0.80 ± 0.20, p < 0.05) were lower in SH than in C lambs. During exercise, glue (SH: 3.59 ± 0.19 vs. C: 5.15 ± 0.80, p < 0.05) and FFA (SH: 0.79 ± 0.32 vs. C: 1.23 ± 0.43, p < 0.05) increased significantly in SH and C lambs (p < 0.05). However, the relative increment of glue during exercise was lower in SH than in C lambs (7 ± 5 % vs. 15 ± 7 %, p < 0.05). The relative increment of FFA was not different (SH: 38 ± 37 % vs. C: 56 ± 47 %, p = 0.48). We conclude that, despite lower gluc and FFA at rest, SH lambs demonstrate a metabolic response of increment of gluc and FFA during moderate exercise like C lambs, However, the relative increment for gluc was lower in SH lambs. We speculate that this is due to an earlier glycogen depleted state in SH lambs.

Effect of infusing branched-chain amino acid during incremental exercise with reduced muscle glycogen content

The aim of this study was to investigate whether, when muscle glycogen is reduced, a pre-exercise infusion of branched-chain amino acids (BCAA) modifies exercise performance or the metabolic and respiratory responses to incremental exercise. Six moderately trained volunteers took part in the following protocol on two occasions. On day 1, at 9 a.m. in the postabsorptive state, they performed a graded incremental exercise (increases of 35 W every 4 min) to exhaustion (Ex-1). A meal of 1,000 kcal (4,200 kJ; 60% protein, 40% fat) was consumed at 12 p.m. No food was then allowed until the end of the experiment (20-21 h later). A 90-min period of exercise at alternating high and moderate intensities, designed to deplete muscle glycogen, was performed between 6 p.m. and 7.30 p.m. The morning after (day 2), the subjects randomly received either a mixed solution of BCAA (260 mg x kg-1 x h-1 for 70 min), or saline. They then repeated the graded incremental exercise to exhaustion (Ex-2). Metabolic and respiratory measurements suggested a muscle glycogen-depleted state had been achieved. No significant differences were observed in total work performed, maximal oxygen uptake or plasma ammonia, alanine, and blood pyruvate concentrations in the two treatments. After BCAA infusion, higher blood lactate concentrations were observed at maximal power output in comparison with those during saline [BCAA 4.97 (SEM 0.41) mmol x l-1, Saline 3.88 (SEM 0.47) mmol x l-1, P < 0.05].(ABSTRACT TRUNCATED AT 250 WORDS)

Hypoglycemia but not hyperglycemia induces rapid changes in pancreatic beta-cell gene transcription

The purpose of these studies was to quantify several mRNAs expressed specifically in pancreatic islet cells and known or postulated to be important for insulin release after acute well defined alterations in levels of plasma glucose. Glucose levels were maintained at 50, 120, or 180 mg/dl (2.8, 6.7, or 10 mM) for 3 h in conscious unrestrained rats. Hypoglycemia (for 3 h) caused significant decreases in pancreatic content of mRNAs for insulin 2 and GLUT-2 to 55 and 34% of control values, respectively. There were no significant changes in insulin 1, amylin, glucokinase, or glucagon mRNAs. Unprocessed insulin 1 and 2 mRNA precursors were decreased to 17 and 10% of levels in controls, consistent with effects of short-term hypoglycemia on new mRNA synthesis. Hyperglycemia (for 3 h) caused no increase in pancreatic content of any mRNA measured. To discriminate between effects of hypoglycemia and hyperinsulinemia in the hypoglycemic animals, rats were made hypoglycemic by infusion with etomoxir, a carnitine palmitoyltransferase I inhibitor that lowers glucose in the fasted (glycogen-depleted) state by inhibiting hepatic gluconeogenesis. A single dose of this agent caused a decrease in glucose from 120 mg/dl (6.7 mM) to 80 mg/dl (4.4 mM) and significantly decreased insulin mRNA and pre-mRNA. These results are consistent with the hypothesis that glucose modulates islet cell gene transcription directly. They indicate that the range of glucose concentrations that modulate gene transcription differs from the levels of glucose that alter both insulin biosynthetic and secretion rates.

Plasma catecholamine and lactate response during graded exercise with varied glycogen conditions

The relationships between the lactate threshold (TLa), plasma catecholamines, and ventilatory threshold (TVE) were examined under normal and glycogen-depleted conditions. Nine male subjects performed a graded exercise test on a bicycle ergometer in a normal glycogen (NG) state and in a glycogen-depleted (GD) state to determine if manipulation of muscle glycogen content would affect their ventilatory, lactate, and catecholamine responses. High correlations were found between plasma lactate and the two catecholamines, epinephrine (r = 0.964) and norepinephrine (r = 0.965) under both conditions. The GD protocol resulted in a shift in the TLa to a later work rate; inflections in epinephrine and norepinephrine shifted in a coordinated manner. TVE and TLa occurred at similar work loads under NG conditions [67.2 +/- 1.5 and 65.6 +/- 2.3% maximal oxygen consumption (VO2max), respectively], but TLa occurred at a later work load (75.3 +/- 1.9% VO2max) compared with TVE (68.3 +/- 1.6% VO2max) under GD conditions. These results suggest a causal relationship between plasma lactate and epinephrine during a graded exercise test under the glycogen conditions studied. Although an association existed between ventilation and lactate, this relationship was not as strong.

Blood glucose turnover during high- and low-intensity exercise.

We hypothesized that whole body glucose uptake (Rd) during exercise is not related in a simple, linear manner to O2 uptake (VO2). To test this, seven healthy male subjects (age range 23-34 yr) were studied in the postabsorptive but not glycogen-depleted state. Three conditions were examined: 1) rest, 2) 40 min of constant exercise in which the work rates were carefully chosen to consist of low-intensity exercise (no elevated blood lactate, a mean of 40% maximal VO2), and 3) 40 min of high-intensity exercise (markedly elevated blood lactate, 79% maximal VO2). Gas exchange was measured breath by breath, and glucose uptake and production were measured using [6,6-2H2]glucose. Low-intensity exercise (n = 7) resulted in a small but not statistically significant increase in mean Rd [3.06 +/- 0.37 (SE) mg.min-1.kg-1] compared with resting values (2.87 +/- 0.39 mg.min-1.kg-1) despite a fourfold increase in the production of CO2 and VO2. By contrast, the high-intensity exercise Rd (n = 5, 6.98 +/- 0.67 mg.min-1.kg-1) was significantly greater than the resting value (3.03 +/- 0.56 mg.min-1.kg-1). Results of glucose production were virtually the same. Similarly, mean levels of epinephrine and norepinephrine increased significantly above resting values during high- but not low-intensity exercise. Our data demonstrate that whole body glucose dynamics and regulation during 40 min of exercise do not change in a simple linear manner with respect to metabolic rate.

The effect of body build, diet and endocrine factors on the extrinsic fibrinolytic system in healthy young women.

In this study, the importance of anthropometric, nutritional and endocrine variables on the plasma concentrations of tissue plasminogen activator antigen (tPA) and plasminogen activator inhibitor (PAI-l) were investigated. Tissue plasminogen activator concentration and PAI-l activity in plasma were studied in 24 healthy young women after diet periods which caused depletion or filling of hepatic glycogen stores. Plasminogen activator inhibitor levels in the glycogen-depleted state and the glycogen-repleted state were positively correlated, as were also tPA levels. In fasting subjects with repleted glycogen stores, tPA values correlated with fasting insulin concentration and blood pressures. In fasting subjects depleted of glycogen stores, PAI-l correlated with tPA, plasma insulin, triglycerides, and waist-to-hip girth ratio; triglycerides and waist-to-hip girth ratio also correlated with tPA. Over a 4-h period following intake of a test-meal, the glucose and insulin responses were not correlated with the fibrinolytic variables. Multivariate regression analysis suggested that waist-to-hip girth ratio and diastolic blood pressure were independently associated with tPA concentrations both in subjects with depleted and repleted glycogen stores. Thus, both constitutional factors such as anthropometric variables and blood pressure, and nutritional status of the subjects may be related to tPA and PAI- levels in plasma. This should be taken into account in clinical studies on fibrinolysis.

Effects of glycogen depletion and pedaling speed on "anaerobic threshold".

Nine male subjects performed continuous incremental exercise on a bicycle ergometer pedaling at 50 and 90 rpm in a normal glycogen state (NG) and at 50 rpm in a glycogen-depleted state (GD) to determine if alterations in pedaling frequency and muscle glycogen content would affect their "anaerobic thresholds." Ventilatory [T(vent)] and lactate [T(lac)] thresholds were identified as the points after which expired minute volume and blood lactate began to increase nonlinearly as a function of work rate. The GD protocol elicited a significant divergence between the two thresholds shifting the T(vent) to a lesser and the T(lac) to a greater work rate relative to the NG state. When the pedaling frequency was increased to 90 rpm in the NG condition, the T(lac) was shifted to a lesser work rate relative to the 50-rpm NG condition. A correlation of only 0.71 was obtained between subjects' T(vent) and T(lac). In subjects of less than 70 kg body wt, the T(lac) came at a work rate 400 kg.m.min-1 less than in subjects of greater than 80 kg body wt despite equivalent O2 uptake. The observation that the T(vent) and T(lac) could be manipulated independently of each other reveals limitations in using the T(vent) to estimate the so-called anaerobic threshold.